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. 2021 Jun 23;7(26):eabg8139. doi: 10.1126/sciadv.abg8139

Fig. 3. AC-secreted CCL2 significantly increases the extravasation of TCs via their CCR2 receptor.

Fig. 3

(A) Extravasation efficiency of MDA-231 TCs in triculture MVNs with neutralizing antibodies at 6 hours after perfusion. Abs, antibodies. (B) Representative CCL2 staining in triculture MVNs and quantification of the CCL2 MFI per cell volume in iPS-ECs, PCs, and ACs. (C) Extravasation efficiency of MDA-231 in triculture MVNs with ACs treated with CCL2 small interfering RNA (siRNA) or control siRNA before device seeding. (D) Representative images of MDA-231 in triculture MVNs stained with anti-CD31 at 6 hours after perfusion, in the case of MVNs seeded with ACs treated with CCL2 siRNA or control siRNA. (E) Extravasation efficiency of MDA-231 in triculture MVNs treated with anti-CCR2 and/or anti-CCR4, and the appropriate control at three time points. Significance is assessed with respect to the control [dimethyl sulfoxide (DMSO)] condition. (F) Extravasation efficiency of MDA-231 in triculture MVNs, in the case of MDA-231 treated with anti-CCR2 and/or anti-CCR4, and the appropriate control at three time points. Significance is assessed with respect to the control (DMSO) condition. (G) Representative images of MDA-231 in triculture MVNs stained with anti-CD31, in the case of TCs treated with anti-CCR2 or the appropriate control at 6 hours after perfusion. (H) Representative images of A549 in triculture MVNs stained with anti-CD31, in the case of TCs treated with anti-CCR2 or the appropriate controlat 6 hours after perfusion. (I) Extravasation efficiency of A549 in triculture MVNs, in the case of A549 cells treated with anti-CCR2 and/or anti-CCR4, and the appropriate control at three time points. Significance is assessed with respect to the control (DMSO) condition. (J) Extravasation fold of all four TCs between iPS-EC + AC MVNs and the monoculture iPS-EC MVNs at 24 hours (left) and relative CCR2 gene expression [normalized by glyceraldehyde phosphate dehydrogenase (GAPDH)] for all four TCs measured via qRT-PCR (right). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001; not significant; n.s.#, not significant across all conditions (pairwise).